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Causal Agents of Mycobacterial Diseases in Freshwater Ornamental Fish and Their Importance for Human Health in the Czech Republic

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Causal Agents of Mycobacterial Diseases in Freshwater Ornamental Fish and Their Importance for Human Health in the Czech Republic ACTA VET. BRNO 2006, 75: 251–258 Causal Agents of Mycobacterial Diseases in Freshwater Ornamental Fish and their Importance for Human Health in the Czech Republic J. ¤EHULKA1, J. KAUSTOVÁ2, E. ¤EHULKOVÁ3 1 Department of Zoology, Silesian Museum, Opava, 2 National Reference Laboratory for Mycobacterium kansasii, Regional Institute of Public Health, Ostrava, 3 Department of Zoology and Ecology, Faculty of Science, Masaryk University, Brno, Czech Republic Dedicated to the 80th birthday of Prof. MVDr. Z. Luck˘, CSc., Professor Emeritus of the Department of Game, Fish and Bee Diseases, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic Received September 15, 2005 Accepted March 16, 2006 Abstract ¤ehulka J., J. Kaustová, E. ¤ehulková: Causal Agents of Mycobacterial Diseases in Freshwater Ornamental Fish and their Importance for Human Health in the Czech Republic. Acta Vet. Brno 2006, 75: 251-258. In the period from 1995 to 2004, the following mycobacterial species were isolated during the examination of the state of health of aquarium fish from the Moravian-Silesian region of the Czech Republic: Mycobacterium fortuitum (Paracheirodon axelrodi, Poecilia sphenops, Hemigrammus rhodostomus, Hyphessobrycon flammeus), M. chelonae (P. axelrodi, Betta splendens, Xiphophorus helleri, Puntius tetrazona), M. kansasii (P. axelrodi, B. splendens), M. simiae (Hyphessobrycon ecuadoriensis), and M. gordonae (P. axelrodi, Colisa lalia, B. splendens, Symphysodon discus). The first isolation from aquarium fish was recorded in the case of M. kansasii and the second isolation in the case of M. simiae; both should also be taken into account as aetiological agents of mycobacterial infections in humans. The measures to reduce mycobacterial infections in both fish and humans should include monitoring water mains as possible sources of M. kansasii infections, compliance with the sanitation principles in handling fish suspected of being infected and the content of tanks (molluscs, water, plants), and the introduction of strict veterinary inspection of imported fish, fish from aquarist shops and from large-scale distributors. Zoonosis, mycobacteriosis,“fish-tank” granuloma, Mycobacterium fortuitum, M. chelonae, M. kansasii, M. simiae, M. gordonae Identification of the causal agents of bacterial diseases in aquarium culture is an important part of the health care of ornamental fish, aimed at reducing infections and minimising losses. This applies, in particular, to conditions where ornamental fish are reproduced on a mass scale and therefore, where the probability of an outbreak of epizooties of bacterial aetiology is very high. In addition to the much feared causal agents of acute bacterial diseases, attention must also be paid to the agents causing chronic conditions, including, in particular, mycobacterial infections. According to Mátlová et al. (1998), there are more than 70 (and by the latest data already more than 90) known mycobacterial species that can be classified by their relation to the host into obligate pathogens of vertebrates (Mycobacterium tuberculosis, M. paratuberculosis, M. bovis) and potentially pathogenic mycobacteria, which include the majority of mycobacteria potentially pathogenic for the poikilotermic animals (M. marinum, M. chelonae and M. fortuitum). The first case of mycobacteriosis in freshwater fish was described by Bataillon et al. (1987) in the common carp (Cyprinus carpio): the authors named the isolated bacteria M. piscium. According to literature, mycobacterioses belong to the most common chronic diseases of both freshwater and marine fish all over the world, particularly in the regions of Address for correspondence: Doc. Ing. J. ¤ehulka, CSc. Phone: +420 553 628 660 Slezské zemské muzeum Fax: +420 553 622 999 NádraÏní okruh 33 E-mail: [email protected] 746 01 Opava, Czech Republic http://www.vfu.cz/acta-vet/actavet.htm 252 the temperate and tropical zone. Ippen (1964) states that mycobacterial infections have been described to occur in 10 orders, 85 genera and 123 species of freshwater and marine fish, and according to Nigrelli and Vogel (1963) they have been described in 151 fish of 34 genera from different geographical regions. However, the occurrence of mycobacterial infections is most common in aquarium fish, where they rank among the most dangerous health problems with unfavourable prognosis. Jahnel (1940ab) described findings of mycobacteria in the following species: black tetra (Gymnocorymbus ternetzi), pearl danio (Danio albolineatus), spottedsail barb (Puntius phutunio), malabar danio (Devario malabaricus), Egyptian mouth-brooder (Pseudocrenilabrus multicolor multicolor), bloodfin tetra (Aphyocharax anisitsi), dwarf gourami (Colisa lalia) and representatives of the genus Xiphophorus. Although it is impossible to eliminate a disease entirely in its acute form, when fish dies with no external symptoms, systemic diseases regularly occur, affecting any organ with a typical granulomatous response in the advanced stages of the condition (Ippen 1964; Wolke 1975; Wolke and Stroud 1978; Majeed et al. 1981; Csaba et al. 1982; Noga et al. 1990). Much attention was paid to mycobacterioses by Amlacher (1986). In his monograph about fish diseases, he states that 71 species of aquarium fishes are susceptible, and provides documentation concerning macro- and microscopic findings in 9 species of ornamental fishes. The causative organism of mycobacteriosis in the goldfish (Carassius auratus) and redeye tetra (Moenkhausia sanctaefilomenae) in Malaysia was characterised on the basis of examining its biochemical and physiological properties. The isolate differed in many characteristics from the reference strains of M. chelonae subsp. chelonae and M. chelonae subsp. abscessus as well as the rapidly growing photochromogenic M. parafortuitum and M. vaccae (Shamsudin et al. 1990). As for the species of more recent description, mention should be made of M. poriferae in the snakehead murrel (Channa striatus) (Tortoli et al. 1996). Skin ulcers and fin damage due to mycobacterial infection have been previously reported in aquarium fish (Richards 1977; van Duijn 1981). Not even some salmonid fishes have escaped mycobacterial infections. Wood and Ordal (1958) reported a very high incidence of mycobacterioses in hatchery-reared Pacific salmon and steelhead trout fed ground, uncooked viscera of adult fish infected with mycobacteria. Serology of M. chelonae isolated from salmonids (Salmo clarkii, Oncorhynchus kisutch, O. tshawytscha, Prosopium williamsoni) was performed by Arakawa et al. (1986). Interestingly, cutaneous mycobacteriosis was found in the common carp (Majeed and Gopinath 1983) and in yellow perch ( Perca flavescens) (Daoust et al. 1989). On the Czech territory, the first cases of mycobacterium infection were described by Luck˘ and Vanûk (1962) in the following aquarium fishes: Pearl gourami (Trichogaster leeri), blue gourami (T.trichopterus), green swordtail (Xiphophorus helleri), guppy (Poecilia reticulata), siamese fighting fish (Betta splendens), Agassiz’ dwarf cichlid (Apistogramma agassizii) and giant gourami (Osphronemus gouramy). Increasing occurrence of mycobacterial diseases in aquarium culture encourages the efforts of ichthyopathologists to detect the infectious agent and to enhance the knowledge of the sources of mycobacteria other than tuberculosis (MOTT). This is because the outcomes of experimental work indicate that water as a reservoir of certain mycobacterial species plays an important role in the epidemiology of mycobacterial infections caused by MOTT in people and animals (Eilertsen 1969; ·losárek and Janota 1990). The objective of the study was to provide information on current knowledge of the clinically important mycobacterial infections in aquarium fish from the Moravian/Silesian region of the Czech Republic with respect to the current growing interest in the issues of the occurrence of mycobacteria in poikilothermic animals and humans (Mátlová et al. 1998). 253 Materials and Methods Fish Over the past decade, as many as 186 freshwater ornamental fish of 74 species have been delivered to our laboratory for an examination of the cause of disease. Most of the fish came from small breeders and from larger reproduction facilities, but some also came from aquarium fish shops and hobby breeders. Post mortem examinations were carried out according to Luck˘ (1982), Roberts and Shepherd (1997), Austin and Austin (1987), Ergens and Lom (1970), and Lom and Dyková (1992). Bacteriology The bacteriological procedures involved Gram staining of tissue smears, cultivation on growth media and evaluation of microbial cultures. Blood agar (Columbia Agar Base MERCK), cytophaga agar (Anacker and Ordal 1959), tryptone soya agar (TSA, Oxoid) and sweet wort agar (Fassatiová 1979) served for the cultivation. Mycobacteriological examination The samples were decontaminated by modified Petroff method with 1M NaOH solution. From the sediment two smears were prepared for Ziehl-Neelsen and fluorochrome staining. Sediment was inoculated onto 3 Löwenstein Jensen media. Two of them were incubated at 37 °C and one at 30 °C. Where acid-fast bacilli (AFB) were found, the strain was identified either by means of the GENProbe chosen on the basis of macro- and microscopic morphology of the strain, or by conventional methods based on growth characteristics: temperature tolerance, colony and cell morphology, growth rate, pigment production and biochemical properties. Susceptibility to streptomycin (STM), isoniazide (INH),
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